LOCUS       sequence-255132-        7648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac transposon vector that Inducibly (Tet-On) expresses RTP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7648)
  AUTHORS   Jones EM, Jajoo R, Cancilla D, Lubock NB, Wang J, Satyadi M, Cheung 
            R, de March C, Bloom JS, Matsunami H, Kosuri S
  TITLE     A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions 
            with RNA Sequencing.
  JOURNAL   Cell Syst. 2019 Mar 27;8(3):254-260.e6. doi: 
            10.1016/j.cels.2019.02.009. Epub 2019 Mar 20.
  PUBMED    30904378
REFERENCE   2  (bases 1 to 7648)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cels.2019.02.009"; journalName: "Cell Syst"; date: 
            "2019-03-27- 27"; volume: "8"; issue: "3"; pages: "254-260.e6"
COMMENT     Sequence Label: pB-RTP2-Integration
FEATURES             Location/Qualifiers
     source          1..7648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1406..1424
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1443..1461
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1480..1498
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1517..1535
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1554..1572
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1591..1609
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1628..1646
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1665..1683
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1705..1729
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    2733..2867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2978..3189
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3202..3470
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3497..4096
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3497..3516)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3993..4013
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4156..4176)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4218..4339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   5102..5136
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5627..5643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5627..5643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5640..5662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5651..5667
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5675..5681,5682..5699,5700..5705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5720..5741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5858..5875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6029..6617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6109..6128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6788..7579,7580..7648))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7411..7430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtccaatcat aggggtaccc
     1381 gacccatcaa gtgccacctg acgtctccct atcagtgata gagaagtcga cacgtctcga
     1441 gctccctatc agtgatagag aaggtacgtc tagaacgtct ccctatcagt gatagagaag
     1501 tcgacacgtc tcgagctccc tatcagtgat agagaaggta cgtctagaac gtctccctat
     1561 cagtgataga gaagtcgaca cgtctcgagc tccctatcag tgatagagaa ggtacgtcta
     1621 gaacgtctcc ctatcagtga tagagaagtc gacacgtctc gagctcccta tcagtgatag
     1681 agaaggtacc ccctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
     1741 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tgagctcgtt
     1801 tagtgaccgt ctagagccgc caccatgtgt acttcactga ccacgtgtga gtggaagaag
     1861 gtgttttatg agaaaatgga agtcgccaag cctgccgatt cctgggaact tatcatcgac
     1921 ccaaacctga agccatccga gctcgctccc gggtggaagc agtacctgga acaacacgct
     1981 tccggaaggt tccactgctc ttggtgctgg cacacttggc agtcagcaca tgtagtcatt
     2041 ctgttccaca tgtttctgga tagggctcaa cgggcaggca gcgtgcgcat gcgcgtcttt
     2101 aagcaactct gctacgagtg tggcacagcc cgacttgatg agtcttcaat gttggaagag
     2161 aacatcgagg gcctggtgga caatctcatc acgtctttgc gcgaacagtg ttacgaagag
     2221 gatggggggc aataccggat tcatgttgca tctcgccctg attcaggtcc acacagggca
     2281 gagttctgcg aggcatgtca ggaaggtatt gtgcattgga aaccctccga gaagctgctg
     2341 gaggaggagg ttacaacata tactagcgaa gcgtccaaac ctagagcaca agccgggagc
     2401 ggctacaact tcctttccct tcggtggtgc ctgttttggg cctcactgtg cctgctggtt
     2461 gtgtatttgc agttcagctt cttgagccct gccttttttt gagaattcgc cacacaggca
     2521 tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg
     2581 taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc
     2641 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2701 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2761 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2821 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcccc aggaagctcc
     2881 tctgtgtcct cataaaccct aacctcctct acttgagagg acattccaat catagggcgg
     2941 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     3001 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc
     3061 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     3121 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     3181 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc
     3241 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc
     3301 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg
     3361 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt
     3421 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga
     3481 ccggcgccta cgctagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3541 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3601 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3661 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3721 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3781 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3841 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3901 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3961 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4021 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4081 caagcccggt gcctgagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4141 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4201 gtatcatgcg ttaactaaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4261 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4321 aactcatcaa tgtatcttat catgtctgga attgactcaa atgatgtcaa ttagtctatc
     4381 agaagctcat ctggtctccc ttccggggga caagacatcc ctgtttaata tttaaacagc
     4441 agtgttccca aactgggttc ttatatccct tgctctggtc aaccaggttg cagggtttcc
     4501 tgtcctcaca ggaacgaagt ccctaaagaa acagtggcag ccaggtttag ccccggaatt
     4561 gactggattc cttttttagg gcccattggt atggcttttt ccccgtatcc ccccaggtgt
     4621 ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc
     4681 cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg cggggggagc gccggaccgg
     4741 agcggagccc cgggcggctc gctgctgccc cctagcgggg gagggacgta attacatccc
     4801 tgggggcttt gggggggggc tgtccctgat atctataaca agaaaatata tatataataa
     4861 gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa
     4921 agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc
     4981 agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg
     5041 tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat
     5101 gcatgcgtca attttacgca gactatcttt ctagggttaa tctagctgca tcaggatcat
     5161 atcgtcgggt cttttttccg gctcagtcat cgcccaagct ggcgctatct gggcatcggg
     5221 gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga gtttgccgag
     5281 gatgactgct gctgcattga cgttgagcga aaacgcacgt ttaccatgat gattcgggaa
     5341 ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc aggccacctg ggataccagt
     5401 tcgtcgcggc ttttccggac acagttccgg atggtcagcc cgaagcgcat cagcaacccg
     5461 aacaataccg gcgacagccg gaactgccgt gccggtgtgc agattaatga cagcggtgcg
     5521 gcgctgggat attacgtcag cgaggacggg tatcctggct ggatgccgca gaaatggaca
     5581 tggatacccc gtgagttacc cggcgggcgc gcttggcgta atcatggtca tagctgtttc
     5641 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5701 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5761 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5821 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5881 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5941 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6901 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7081 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7261 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7621 gggcgacacg gaaatgttga atactcat
//
